Allergic asthma is one of the most common phenotypes of asthma, affecting over 330 million people worldwide. The aim of this study is to explore the pathogenesis of allergic asthma by establishing an in vitro model of allergic asthma. Firstly, a microfluidic chip model for studying human epithelial-dendritic cell co-culture has been constructed by combining microfluidics and electrospinning technologies. Then, by utilizing electrospinning technology, a PLGA basement membrane was constructed, and non-contact co-culture of BEAS-2B epithelial cells and dendritic cells on the microfluidic chip was achieved based on this basement membrane. IL-25 and IL-33, as two major pro-inflammatory factors of asthma, were simultaneously detected by combining microfluidic technology and digital ELISA, through the design of optical path. The experimental results showed that the concentrations of IL-25 and IL-33 increased after the addition of dust mite antigen and tended to normalize after the addition of therapeutic drugs. Therefore, the in vitro co-culture microfluidic chip model designed in this study has revealed the pathogenesis of allergic asthma. Moreover, the drug experiments provide important references for future clinical diagnosis and targeted drug therapy.
Indoor dust mites are one of the common and major inhalant allergens, and detection of specific antibodies against dust mite allergens can help in the prevention and treatment of allergic diseases and in the study of immunotherapy. However, traditional assays such as enzyme-linked immunosorbent assays are time-consuming and costly, and are not conducive to rapid detection. Microfluidic chips enable precise control and manipulation of tiny fluids, allowing for rapid, low-cost, high-throughput bioassays. Digital enzyme-linked immunosorbent assays have the advantages of high sensitivity and specificity. In this paper, fluorescence-encoded micro-magnetic beads were used as solid-phase carriers to modify Der p1 dust mite antigen, and then combined with patient serum and detection antibody to form an immunosandwich structure. Then, we successfully detected Der p1 dust mite allergen-specific antibodies in patient sera using a combination of microfluidic chip and digital ELISA, and plotted the standard curve. The method has promising applications and can provide an essential reference for clinical diagnosis and treatment.
Cell migration assay is the most common research approach for cell migration. Quantitative research and analysis are carried out by measuring the migration of cells into the region that is artificially created among confluent monolayer cells. To improve the efficiency and accuracy of the analysis, the software/tools were developed to assist the image analysis process. However, these software and tools are still at the stage of measuring a single sample, which cannot satisfy the requirement of large sample size for cell migration assay device. In this paper, an image analysis tool based on Fiji is developed, which can segment multiple samples from a scanned image and then analyze a single sample in batch. In addition, the screening function should be added for the application scenario of large sample size. The samples can be filtered according to different conditions to improve the consistency of experimental conditions. The results show that the developed analysis tool ATCA has high accuracy in identifying cell-free zones, with a difference of 2.3% from the tool WHST and 2.9% from manual operation. The analysis efficiency of this tool is 15 times that of manual operation.
Uremia is a serious disease in the stage of renal failure, so it is very important to detect uremia in time. In order to overcome the shortcomings of insufficient sensitivity of traditional ELISA methods, we report a digital ELISA detection platform composed of three parts of microsphere operating system, optical imaging system and computer processing system, which can realize the automatic detection of biomarkers in serum. The platform can combine fluorescent coding microspheres with microporous array, and apply DEP electric field externally, which can effectively increase the recognition efficiency of fluorescent coding microspheres and realize the ultra-high sensitivity detection of two important cytokines of IL-6 and IL-4 in uremic serum. The experiment successfully and accurately detected the IL-4 and IL-6 in the serum of uremic patients. At the end of the article, we summarize and prospect the experiment.
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